摘要
目的探讨自噬在卵巢癌失巢凋亡抵抗中的作用。分析自噬和整合素相关蛋白在卵巢癌细胞株A2780不同培养方式下的表达情况,并通过siRNA和化学抑制剂来明确其调控机制。方法 2012年6月至2013年6月在上海交通大学医学院附属仁济医院以卵巢癌细胞株A2780在常规培养、Matrigel铺胶培养和悬浮培养这3种培养方式中用Western blot检测自噬和整合素相关蛋白的表达,流式细胞技术检测细胞凋亡情况。结果卵巢癌细胞株A2780在常规培养、Matrigel铺胶培养和悬浮培养这3种培养方式中自噬相关分子LC3、Beclin1和整合素CD51(αv),CD61(β3),CD29(β1)的表达情况发现,CD51和CD61的表达逐渐下降(P<0.05),而自噬相关分子LC3的表达却为递增(P<0.05),Beclin1蛋白表达差异无统计学意义(P>0.05)。在3种不同培养方式中,RGD使用后自噬相关分子LC3的蛋白表达增强(P<0.05);RGD阻断整合素信号途径并没有促进细胞的凋亡(P>0.05),但是RGD作用下同时加入自噬抑制剂3-MA可以促进细胞的凋亡(P<0.05)。结论自噬在卵巢癌失巢凋亡中主要通过整合素信号αvβ3途径来调控,可以为失巢细胞早期提供能量支持,在一定条件下抵抗失巢凋亡,这也是造成卵巢恶性肿瘤转移的重要原因。
Objective To explore the role of autophagy in anoikis resistance for ovarian cancer cells as well its association with apoptosis. Methods Between June 2012 and June 2013 in Shanghai Jiaotong University School of Medicine, analysis of autophagy related protein and integrin related protein in A2780 ovarian cancer ceils with different culture condition. siRNA and chemical inhibitor is used to investigate the regulation mechanism, real time PCR and western blot were used to investigate the expression of autophagy and integrin molecular in ovarian cancer cells, cells apoptosis is examined by flow cytometry. Results Comparing among routine culture, matrigel culture and suspend culture, the increasing LC3 ex- pression of autophagy（ P 〈 0.05 ） accompanied with integrin CDS1 （ o^v）, CD61 （ 133 ） decreasing（ P 〈 0.05 ）, but there was no difference in protein expression of Beclinl and CD29 （ β1 ） （ P 〉 0.05 ）. Inhibiting integrin αvβ3 pathway by RGI）, autophagy protein expression of LC3 was increased（P 〈 0.05 ）. PtGI） promoted cell apoptosis with autophagy inhib- itor 3-MA （ P 〈 0.05 ）, but not without 3-MA （ P 〉 0.05 ）. Conclusion Autophagy pathway played a potentially protective role in anoikis apoptosis. Intercellular signaling pathways of integrin might play a regulatory role on autophagy in ovarian cancer anoikis resistance which resulted in distant metastasis of cancers.
出处
《中国实用妇科与产科杂志》
CAS
CSCD
北大核心
2014年第1期46-50,共5页
Chinese Journal of Practical Gynecology and Obstetrics
基金
国家自然科学基金面上项目(81072137)
关键词
自噬
卵巢癌
失巢凋亡
整合素
autophagy
ovarian cancer
anoikis
integrin
